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Enhancing the weaver ant, Oecophylla smaragdina (Hymenoptera: Formicidae), for biological control of a shoot borer, Hypsipyla robusta (Lepidoptera: Pyralidae), in Malaysian mahogany plantations Grace T. Lim Dissertation submitted to the faculty of the Virginia Polytechnic Institute and State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy In Entomology Scott M. Salom, Co-Chair Loke T. Kok, Co-Chair Laurence G. Kirton (FRIM) Douglas G. Pfeiffer Richard D. Fell April 4, 2007 Blacksburg, VA Keywords: Weaver ant, Mahogany Shoot Borer, Oecophylla smaragdina, Hypsipyla robusta, Biological control, Food supplementation, Host plant species Enhancing the weaver ant, Oecophylla smaragdina (Hymenoptera: Formicidae), for biological control of a shoot borer, Hypsipyla robusta (Lepidoptera: Pyralidae), in Malaysian mahogany plantations Grace T. Lim Abstract The weaver ant is a promising biological control agent of a shoot borer, Hypsipyla robusta Moore, on mahogany, but techniques to conserve ant colonies redistributed to mahogany plantations have not yet been developed. The effect of food supplementation and host plant species preference of the weaver ant, Oecophylla smaragdina F., was evaluated in a series of field studies. A simple model was developed to estimate the number of ants within nests on Khaya ivorensis A. Chev. (Meliaceae): log10 (Number of ants) = - 1.16 + 1.09 log10 (Nest size). Nest 2 size is calculated from estimated nest height (h) and length (Î) using the formula = pr Î, where r = ½ h. This model was useful for repeated assessments of ant population levels to evaluate treatment effects. It provides better estimates than previous indirect methods based on nest counts and ant trail counts on plant parts. Colonies that were relocated without their queens and very small colonies (< 10,000 ants) failed to establish on new host trees, indicating that a minimum ant population and queen needs to be transferred for colony survival. Established colonies consumed more high- protein foods (live mealworms and fish) than high-carbohydrate liquid foods (honey and ‘weaver ant formula’, which contained sucrose and human muscle-training powder (EnerproTM)). Relocated colonies consumed more weaver ant formula and as many mealworms as established colonies, indicating that existing and relocated colonies require different food supplementation strategies. Decreasing consumption over time and preferential consumption among high-protein food choices (i.e., of mealworms over fish) indicated that ants select and regulate food consumption based on colony needs. Therefore, food supplementation should be as needed. Preliminary indications were that self-sufficiency in trophobiont (honeydew) levels may be achieved in two months after colony relocation. The optimal colony density that would protect K. ivorensis was estimated to be within the range of 6 – 48 colonies per ha based on previous reports for cocoa and cashew, and a consideration of the low damage threshold for mahogany. Substituting chemical control with weaver ants at those application rates gave similar IRRs (Internal rate of return; 11.6 – 12.2 vs. 12.0%) in preliminary financial analyses, and was preferable from an ecological standpoint. Twenty-nine host plant species were found for Malaysian O. smaragdina, of which 11 were new species records for Oecophylla spp. Also, there were two new genera and eight new species records for Malaysian O. smaragdina. Of eight trophobiont families collected, six species were identified, yielding new trophobiont-host plant species records for four coccoid species and two membracid genera. Screening of several ant-abundant plant species that included preliminary pest risk analyses for trophobionts on K. ivorensis, identified M. citrifolia as a promising candidate for mixed-planting with this mahogany species. Acknowledgments I am greatly indebted to my co-advisors, Drs. Scott Salom, Loke T. Kok and Laurence Kirton, for their constant encouragement and guidance throughout my graduate study. I am also grateful to my other advisory committee members, Drs. Richard Fell and Douglas Pfeiffer for their insightful comments and constructive criticism of my work. I wish to appreciate the faculty of Virginia Tech who contributed to my well-rounded graduate education. In addition to all members of my advisory committee, these were Drs. Donald Mullins, Carlyle Brewster and Jeffrey Bloomquist, and Andy Roberts (Entomology Department), Dan Spitzner, Geoff Vining and George Terrell (Statistics Department). Drs. Brewster and Vining provided valuable statistical advice. I also wish to thank Eric Day, Theresa Dellinger, Sarah Kenley, Ashley Lamb, Geoff Preston, Kathy Shelor and Robin Williams (Entomology Department, Virginia Tech) for their encouragement and technical support. I gratefully acknowledge my fellow entomology graduate students for their friendship and advice: Brian Eisenback, Robbie Flowers, Nathan Herrick, Dave Mausel and Meredith Worthen. Shicai Yan provided Chinese – English translation of an important weaver ant paper. I would like to express my warmest appreciation to the staff of the Entomology Unit, Forest Research Institute Malaysia (FRIM) who assisted in my field work: Saimas Ariffin, Saiful Azahari, Shaiful Amri, Norziah Ishak, Fadir Ishak and Azmi Mahyudin. Thanks also to Shawn Cheng and Nada Badruddin, and again to Dr. Kirton, my colleagues at the Entomology Department, FRIM for holding the fort while I was on study leave. Lim Sun Heng, Leong Lyne Ching and Lim Ban Aik, and the staff of Trolak Plantation: Lim Thaw Nam, Tee Aun Seng and Tan Hong Tick (Maju Aik Ltd.) were involved in the project from the start. Thank you. Additionally, I am grateful to Dr Michael Kosztarab (Entomology Department, Virginia Tech), Drs. Douglass Miller and Stuart MacKamey (USDA-ARS) for identifying various trophobionts associated with the weaver ant. The host plant species of the ant were in part identified by Mohd. Asri and Kamarudin Salleh (Botany Section, FRIM). Thanks also to Drs. Nico Blüthgen (Bayreuth University) and Paul van Mele (CABI) for advice on aspects of weaver ant biology and Drs. Marianne Horak (CSIRO) for information on Hypsipyla taxonomy. I am grateful to FRIM for allowing me to pursue my studies while employed at the Institute and for providing partial funding. My sincere thanks to Drs. Abdul Razak Mohd. Ali, Wan Razali Wan Mohd., Abdul Latif Mohmod, Baskaran Krishnapillay, Abdul Rasip Abdul Ghani and Ms. Wan Rahmah Wan Raof (FRIM). I would also like to extend my gratitude to Liza Ismail and Ruziah Ripin (FRIM) for their technical support. Drs. Gary Theseira, Ahmad Zuhaidi Yahya, Ang Lai Hoe, Saw Leng Guan, Chan Hung Tuck, Lillian Chua and Lee Su See are very much appreciated for their help. Last but not least, I would like to express my appreciation and love to my fiancé Spencer Clark and our families for their tremendous support of my graduate education. This project was sponsored by the Fulbright Program that provided full funding for the first two years, Virginia Tech that granted a teaching assistantship for the third and final year and tuition waivers throughout, the Malaysian-American Commission on Educational Exchange that provided a return airfare journey, and FRIM. iii Table of Contents Page Abstract ii Acknowledgements iii List of Tables vi List of Figures vii Preface x Chapter 1 Introduction and Literature Review 1 Characteristics of mahogany 1 Characteristics of the mahogany shoot borer 2 Mahogany shoot borer management 2 Weaver ants 4 Research Objectives 6 Chapter 2 Host plants and associated trophobionts of the weaver ant Introduction 7 Materials and Methods 8 Literature survey 8 Host plant survey 9 Statistical analyses 11 Results 12 Literature survey 12 Host plant survey 12 Discussion 48 Literature survey 48 Host plant survey 48 Chapter 3 The preference of the weaver ant for four plant species in a mangrove habitat Introduction 51 Materials and Methods 52 Statistical analyses 53 Results 53 Discussion 56 Chapter 4 A model for estimating the number of weaver ants inside nests Introduction 59 Materials and Methods 60 Statistical analyses 61 Results 61 Discussion 68 Chapter 5 The preference of the weaver ant for four foods Introduction 70 Materials and Methods 71 Preliminary study 71 iv Choice and no-choice tests 72 Statistical analyses 74 Results 74 Discussion 80 Chapter 6 The effect of food supplementation on the establishment of redistributed weaver ant colonies Introduction 86 Materials and Methods 87 Statistical analyses 90 Results 90 First week 90 16 weeks 93 Discussion 96 First week 96 16 weeks 97 Chapter 7 Conclusions 102 Model for ant population estimation 102 Harvesting and redistribution 102 Direct food supplementation 102 Indirect food supplementation 103 Further Study 104 Literature Cited 105 Appendix 117 Vita 118 v List of Tables Table Page 2.1. Description of sites sampled in four habitats for Oecophylla smaragdina host 10 plants in Central West Peninsular Malaysia. 2.2. Records of Oecophylla smaragdina host plants and possible host plants, and 13 associated trophobionts from a survey of the literature (1900 to present). Currently accepted species and family names are used followed by names given in the original article within square brackets [ ], where different. 2.3. Records of Oecophylla longinoda host plants and associated trophobionts from 32 a survey of the literature (1900 to present). Currently accepted species and family names are used followed by names given in